A surveillance of enteropathogens in piglets from birth to seven days of age in Brazil / Levantamento dos enteropatógenos de leitões do nascimento a sete dias de idade no Brasil

The purpose of the study was to evaluate the real importance of anaerobic enteropathogens and rotavirus in contrast to more common agents as cause of diarrhea in piglets within the first week of life. Sixty 1- to 7-day-old piglets, 30 diarrheic and 30 non-diarrheic (control), from 15 different herds were selected, euthanized and necropsied. Samples of the jejunum, ileum, colon, cecum and feces were collected from the piglets and analyzed to determine the presence of the following enteropathogens: enterotoxigenic Escherichia coli (ETEC), Clostridium perfringens types A and C, Clostridium difficile, rotavirus and Isospora suis. Among diarrheic piglets, 23.3% were positive for C. difficile, 70% for C. perfringens type A cpb2+, 14.3% for rotavirus and 10% for ETEC. Among non-diarrheic control piglets, 10% were positive for C. difficile, 76.7% for C. perfringens type A cpb2+, 0% for rotavirus, 3.3% for ETEC and 3.3% for I. suis. C. perfringens type C was not detected in any of the animals. Histological lesions characteristic of C. difficile, E. coli and rotavirus were observed. However, no C. perfringens type A suggestive lesions were detected. There was a positive correlation between mesocolon edema and the presence of C. difficile toxins. Although C. perfringens type A cpb2+ was the most frequently detected enteropathogen, there was no association between its presence and diarrhea or macro or microscopic changes. C. difficile and Rotavirus were the most relevant pathogens involved with neonatal diarrhea in this study, and histopathology associated with microbiological test proved to be the key to reach a final diagnosis.(AU)

O objetivo do presente estudo foi avaliar a real importância de enteropatógenos anaeróbios e rotavirus em comparação à outros agentes mais comuns como causa de diarreia em leitões até cinco dias de idade. Leitões com 0 a 7 dias de vida, 30 diarreicos e 30 não diarreicos (controles) de 15 granjas diferentes foram eutanasiados e necropsiados. Amostras de jejuno, íleo, colon e ceco foram coletadas e submetidas à detecção dos seguintes enteropatógenos: Escherichia coli enterotoxigênica (ETEC), Clostridium perfringens, Clostridium difficile, rotavirus e Isospora suis. Entre os animais diarréicos, 23.3% foram positivos para C. difficile, 70% para C. perfringens tipo A cpb2+, 14.3% para rotavirus e 10% para ETEC. Entre os leitões não-diarréicos, 10% foram positivos para C. difficile, 76.7% para C. perfringens tipo A cpb2+, 3.3% para ETEC e 3.3% for I. suis. C. perfringens tipo C não foi detectado em nenhum animal. Lesões histológicas características de C. difficile, E. coli e rotavirus foram observadas. Por outro lado, nenhuma lesão sugestiva de C. perfringens foi detectada. Foi possível observar uma correlação positiva entre edema de mesocolon e presença das toxinas A/B. Apesar de C. perfringens tipo A cpb2+ ter sido o patógeno mais encontrado, nenhuma associação com lesões foi encontrada. C. difficile e Rotavirus foram os agentes mais relevantes associados à diarreia neonatal, e ficou demonstrada a relevância de associação de histopatologia com testes de detecção microbiológica para se firmar um diagnóstico.(AU)

Presença de Clostridium perfringens em frangos de corte provenientes de aviários da região de Ribeirão Preto – SP / Presence of clostridium perfringens in broiler from chicken farms in Ribeirão Preto-SP

Visando estudar a presença de Clostridium perfringens em frangos de cortes provenientes de aviários da região de Ribeirão Preto-SP, amostras de conteúdo cecal de aves foram pesquisadas quanto a presença desse microrganismo. As amostras foram semeadas em meios seletivos para clostrídios e incubadas anaerobica­mente. As culturas foram identificadas e caracterizadas pelo método de Gram e séries bioquímicas. Posteriormente realizou-se teste de inoculação em camundongos para confirmar patogenicidade. De um total de 560 amostras coletadas, 374(66,78%) foram positivas para o gênero Clostridium, sendo 94 (16,78 %) Clostridium perfringens. Verificou–se que 19 (3,4%) amostras foram positivas para outras espécies de clostrídios patogênicos como Clostridium chauvoei (3 - 0,54 %), Clostridium sordelli (9 - 1,61%), Clostridium bifermentans (3 - 0,54%), Clostridium septicum (3 - 0,54%) e Clostridium tetani (1 - 0,18%).

The aim of the present study was to detect Clostridium perfringens in broiler chickens from intensive poultry farms in Ribeirão Preto-SP. We collected 516 samples of caecal content that were cultivated onto selective medium for Clostridium and incubated under anaerobic conditions. Gram smears were carried out from the positive samples and identified using biochemical tests. Mice inoculation test was performed in order to confirm pathogenicity. The results showed that 66.78% of the samples were positive for Clostridia, of which 16.78% were C. perfringens. It was confirmed that 19 (3.4%) of the samples were positive to other Clostridia pathogenic species such as Clostridium chauvoei (3 - 0.54%), Clostridium sordelli (9 - 1.61%), Clostridium bifermentans (3 - 0.54%), Clostridium septicum (3 - 0.54%) and Clostridium tetani (1 - 0.18%).

Presença de Clostridium perfringens em frangos de corte provenientes de aviários da região de Ribeirão Preto SP / Presence of clostridium perfringens in broiler from chicken farms in Ribeirão Preto-SP

Visando estudar a presença de Clostridium perfringens em frangos de cortes provenientes de aviários da região de Ribeirão Preto-SP, amostras de conteúdo cecal de aves foram pesquisadas quanto a presença desse microrganismo. As amostras foram semeadas em meios seletivos para clostrídios e incubadas anaerobica­mente. As culturas foram identificadas e caracterizadas pelo método de Gram e séries bioquímicas. Posteriormente realizou-se teste de inoculação em camundongos para confirmar patogenicidade. De um total de 560 amostras coletadas, 374(66,78%) foram positivas para o gênero Clostridium, sendo 94 (16,78 %) Clostridium perfringens. Verificou–se que 19 (3,4%) amostras foram positivas para outras espécies de clostrídios patogênicos como Clostridium chauvoei (3 - 0,54 %), Clostridium sordelli (9 - 1,61%), Clostridium bifermentans (3 - 0,54%), Clostridium septicum (3 - 0,54%) e Clostridium tetani (1 - 0,18%).(AU)

The aim of the present study was to detect Clostridium perfringens in broiler chickens from intensive poultry farms in Ribeirão Preto-SP. We collected 516 samples of caecal content that were cultivated onto selective medium for Clostridium and incubated under anaerobic conditions. Gram smears were carried out from the positive samples and identified using biochemical tests. Mice inoculation test was performed in order to confirm pathogenicity. The results showed that 66.78% of the samples were positive for Clostridia, of which 16.78% were C. perfringens. It was confirmed that 19 (3.4%) of the samples were positive to other Clostridia pathogenic species such as Clostridium chauvoei (3 - 0.54%), Clostridium sordelli (9 - 1.61%), Clostridium bifermentans (3 - 0.54%), Clostridium septicum (3 - 0.54%) and Clostridium tetani (1 - 0.18%).(AU)

Neonatal diarrhea in piglets associated with cpb-2 positive Clostridium perfringens

Clostridium perfringens type A (CPA) has been recognized as one of the most important cause of neonatal diarrhea in piglets. Despite its importance, the pathogenesis of CPA-associated disease is still unclear and data regarding its occurrence in Brazil is scarce. In light of this, the aim of this study was to report a case of neonatal diarrhea in piglets by CPA encoding beta-2 toxin gene (cpb-2). Three three-day-old piglets from a 2000-sow herd with history of diarrhea were necropsied and intestinal samples were collected for histology,  immunohistochemistry, and feces samples were collected for bacteriologic and molecular procedures. Gross and histopathology revealed superficial necrotic enteritis associated with colonies of bacilli adhered to the exposed lamina propria. These ileal and jejunum fragments were positive for C. perfringens by immunohistochemistry, while anaerobic colonies were identified by PCR multiplex as CPA with the cpb-2. No other enteropathogen was identified from intestinal samples. The C. perfringens isolated strains were susceptible to penicillin, metronidazole and vancomicyn and resistant to eritromycin, enrofloxacin, oxitetracyclin and lincomycin.

Neonatal diarrhea in piglets associated with cpb-2 positive Clostridium perfringens

Clostridium perfringens type A (CPA) has been recognized as one of the most important cause of neonatal diarrhea in piglets. Despite its importance, the pathogenesis of CPA-associated disease is still unclear and data regarding its occurrence in Brazil is scarce. In light of this, the aim of this study was to report a case of neonatal diarrhea in piglets by CPA encoding beta-2 toxin gene (cpb-2). Three three-day-old piglets from a 2000-sow herd with history of diarrhea were necropsied and intestinal samples were collected for histology,  immunohistochemistry, and feces samples were collected for bacteriologic and molecular procedures. Gross and histopathology revealed superficial necrotic enteritis associated with colonies of bacilli adhered to the exposed lamina propria. These ileal and jejunum fragments were positive for C. perfringens by immunohistochemistry, while anaerobic colonies were identified by PCR multiplex as CPA with the cpb-2. No other enteropathogen was identified from intestinal samples. The C. perfringens isolated strains were susceptible to penicillin, metronidazole and vancomicyn and resistant to eritromycin, enrofloxacin, oxitetracyclin and lincomycin.(AU)

Molecular detection and characterization of cpb2 gene in Clostridium perfringens isolates from healthy and diseased chickens

Clostridium perfringens is an important pathogen in both human and veterinary medicine. Necrotic enteritis (NE) is the most clinically dramatic bacterial enteric disease of poultry induced by C. perfringens. The pathogenicity of this bacterium is associated with the production of extracellular toxins produced by some of its strains, such as beta2 toxin. The exact role of beta2 toxin in NE pathogenesis is still controversial. In the present study, C. perfringens isolates from healthy and diseased poultry flocks from different parts of Iran were analyzed by PCR assay to determine the presence of all variants of the beta2 toxin gene (cpb2). The products of two positive cpb2 PCR reactions were sequenced, compared to each other and to the cpb2 sequences published in GenBank (by multiple alignment and phylogenetic analysis). The current work represents the first study of cpb2 in poultry C. perfringens isolates in Asia, and reports the highest percentage of cpb2-positive isolates in both apparently healthy chickens (97.7 percent) and those afflicted with NE (94.4 percent). The sequenced isolates were classified as atypical. This study did not show a direct correlation between NE occurrence and cpb2 presence.(AU)

Molecular detection and characterization of cpb2 gene in Clostridium perfringens isolates from healthy and diseased chickens

Clostridium perfringens is an important pathogen in both human and veterinary medicine. Necrotic enteritis (NE) is the most clinically dramatic bacterial enteric disease of poultry induced by C. perfringens. The pathogenicity of this bacterium is associated with the production of extracellular toxins produced by some of its strains, such as beta2 toxin. The exact role of beta2 toxin in NE pathogenesis is still controversial. In the present study, C. perfringens isolates from healthy and diseased poultry flocks from different parts of Iran were analyzed by PCR assay to determine the presence of all variants of the beta2 toxin gene (cpb2). The products of two positive cpb2 PCR reactions were sequenced, compared to each other and to the cpb2 sequences published in GenBank (by multiple alignment and phylogenetic analysis). The current work represents the first study of cpb2 in poultry C. perfringens isolates in Asia, and reports the highest percentage of cpb2-positive isolates in both apparently healthy chickens (97.7%) and those afflicted with NE (94.4 %). The sequenced isolates were classified as atypical. This study did not show a direct correlation between NE occurrence and cpb2 presence.(AU)

Focal symmetrical encephalomalacia in sheep / Encefalomalacia focal simétrica em ovino

Focal symmetrical encephalomalacia (FSE) is the most prominent lesion seen in the chronic form of enterotoxemia by Clostridium perfringens type D. This paper reports FSE in sheep in Brazil. Six deaths occurred within a seven days period in a flock of 70, four to 30-month-old Santa Inês sheep in the state of Paraíba in the Brazilian semiarid. The flock was grazing a paddock of irrigated sprouting Cynodon dactylon (Tifton grass), and supplemented, ad libitum, with a concentrate of soybean, corn and wheat. Nervous signs included blindness and recumbence. A 19 month-old sheep was examined clinically and necropsied after a clinical course of three days. Gross lesions were herniation of the cerebellar vermis and multifocal, bilateral, symmetric brownish areas in the internal capsule, thalamus and cerebellar peduncles. Histologic lesions were multifocal, bilateral malacia with some neutrophils, swelling of blood vessels endothelium, perivascular edema, and hemorrhages. The flock was vaccinated, before the outbreak, with only one dose of Clostridium perfringens type D vaccine. Two factors are suggested to be important for the occurrence of the disease: insufficient immunity due to the incorrect vaccination; and high nutritional levels by the supplementation with highly fermentable carbohydrates.(AU)

Encefalomalacia focal simétrica (EFS) é a lesão mais proeminente vista nas formas subaguda ou crônica da enterotoxemia por Clostridium perfringens tipo D. Este trabalho relata EFS em ovinos no semiárido do estado da Paraíba. Seis ovinos morreram, em um período de sete dias, dentro de um rebanho de 70 animais, da raça Santa Inês, entre 4-30 meses de idade, que pastavam em piquete de Cynodon dactylon (capim Tifton), que estava rebrotando. Os ovinos eram suplementados com um concentrado de soja, trigo e milho. Os sinais nervosos incluíam cegueira e decúbito lateral. Um ovino de 19 meses de idade foi examinado clinicamente e necropsiado, depois de um curso clínico de 3 dias. Macroscopicamente foram observadas herniação do cerebelo e áreas acastanhadas, multifocais, simétricas e bilaterais na cápsula interna, tálamo e pedúnculo cerebelar. Histologicamente observou-se malacia, bilateral e simétrica, com alguns neutrófilos, tumefação das células endoteliais dos vasos sanguíneos, edema perivascular e hemorragia. O rebanho foi vacinado, antes do surto, com umaúnica dose de vacina para Clostridium perfringens tipo D. Dois fatores são sugestivos quanto a importância da ocorrência da enfermidade: imunidade insuficiente devida à vacinação incorreta; e altos níveis nutricionais da suplementação com carboidratos altamente fermentáveis.(AU)

Clostridium perfringens isolate typing by multiplex PCR

Clostridium perfringens is an important pathogen that provokes numerous different diseases. This bacterium is classified into five different types, each of which capable of causing a different disease. There are various methods for the bacterial identification, many are labor-intensive, time-consuming, expensive and also present low sensitivity and specificity. The aim of this research was to identify the different types of C. perfringens using PCR molecular method. In this study, 130 sheep-dung samples were randomly collected from areas around the city of Kerman, southeastern Iran. After processing and culturing of samples, the produced colonies were morphologically studied, gram stain test was also carried out and the genera of these bacteria were identified through biochemical tests. DNA extracted from isolated bacteria for genotyping was tested by multiplex PCR with specific primers. Based on length of synthesized fragments by PCR, toxin types and bacterial strains were detected. C. perfringens isolated types were divided as follows: 17.39 percent type A, 21.74 percent type B, 34.78 percent type C and 26.09 percent type D. It should be emphasized that, up to the present moment, C. perfringens type A has not been reported in Iran.(AU)

An unusual gangrenous goat mastitis caused by Staphylococcus aureus, Clostridium perfringens and Escherichia coli co-infection / Mastite gangrenosa caprina atípica causada por co–infecção por Staphylococcus aureus, Clostridium perfringens e Escherichia coli

Relata-se, pela primeira vez no Brasil, a ocorrência de mastite gangrenosa caprina atípica causada pela co-infecção por Staphylococcus aureus, Clostridium perfringens e Escherichia coli em uma cabra da raça Boer, na segunda semana de lactação. Descrevem-se os achados clínicos, os procedimentos de diagnóstico microbiológico e a conduta terapêutica.(AU)